New sustainable sources of feeds, which can enhance the health and welfare of farm animals, lower feeding costs, and lead to safer products, are of high priority in modern animal production systems. In the present study, a novel silage created from Greek olive, winery, and feta cheese waste by-products, was evaluated as a feed ingredient at different inclusion rates (0%, 5% or 10%) in 34-day-old weaned pigs. The potential beneficial effects on performance, health and intestinal digesta microflora balance of the pigs were evaluated. Additionally, chemical, microbiological and quality analysis of the meat was carried out. Results showed no detrimental effects (p > 0.05) on the pigs’ performance and no significant changes (p > 0.05) in meat pH, color and chemical analysis. Ileum and cecum microflora populations (total anaerobes, Lactobacillaceae) were positively affected (p ≤ 0.05) by the dietary usage of the silage. The microbial populations (Clostridium spp.) of belly meat cuts were positively modified (p ≤ 0.01). The concentration of total phenols in the meat cuts were increased (p ≤ 0.05) and their resistance to oxidation was improved (p ≤ 0.05). In addition, the fatty acid profile of the meat lipids (polyunsaturated and n-3 fatty acids) was positively modified (p ≤ 0.001).

Antimicrobial resistance is one of the drastically increasing major global health threats due to the misuse and overuse of antibiotics as traditional antimicrobial agents, which render urgent the need for alternative and safer antimicrobial agents, such as essential oils (EOs). Although the strong antimicrobial activity of various EOs has already been studied and revealed, their characteristic high sensitivity and volatility drives the need towards a more efficient drug administration method via a biomaterial system. Herein, the potential of Thymus sibthorpii EO incorporated in functionalized antibacterial collagen hydrogels was investigated. At first, the optimally stabilized type I collagen hydrogels via six different multi-arm poly (ethylene glycol) succinimidyl glutarate (starPEG) crosslinkers were determined by assessing the free amine content and the resistance to enzymatic degradation. Subsequently, 0.5, 1, and 2% v/v of EO were incorporated into optimized collagen hydrogels, and the release profile, as well as release kinetics, were studied. Finally, biomaterial cytocompatibility tests were performed. Thymus sibthorpii EO was released from the hydrogel matrix via Fickian diffusion and showed sustained release and 0.5% v/v EO-loaded hydrogels showed adequate antibacterial activity against Staphylococcus aureus and did not show any statistically significant difference compared to penicillin (p < 0.05). Moreover, none of the fabricated composite antibacterial scaffolds displayed any cytotoxicity on NIH-3T3 fibroblasts. In conclusion, this work presents an innovative antibacterial biomaterial system for tissue engineering applications, which could serve as a promising alternative to antibiotics, contributing to coping with the issue of antimicrobial resistance.

Milk’s biological origin determination, including its adulteration and authenticity, presents serious limitations, highlighting the need for innovative advanced solutions. The utilisation of proteomic technologies combined with personalised algorithms creates great potential for a more comprehensive approach to analysing milk samples effectively. The current study presents an innovative approach utilising proteomics and neural networks to classify and distinguish bovine, ovine and caprine milk samples by employing advanced machine learning techniques; we developed a precise and reliable model capable of distinguishing the unique mass spectral signatures associated with each species. Our dataset includes a diverse range of mass spectra collected from milk samples after MALDI-TOF MS (Matrix-assisted laser desorption/ionization-time of flight mass spectrometry) analysis, which were used to train, validate, and test the neural network model. The results indicate a high level of accuracy in species identification, underscoring the model’s potential applications in dairy product authentication, quality assurance, and food safety. The current research offers a significant contribution to agricultural science, providing a cutting-edge method for species-specific classification through mass spectrometry. The dataset comprises 648, 1554, and 2392 spectra, represented by 16,018, 38,394, and 55,055 eight-dimensional vectors from bovine, caprine, and ovine milk, respectively.

In modern animal production systems, the use of alternative feed sources is essential for reducing production costs, protecting natural resources, and improving meat quality. This study evaluated the impact of a unique silage—one produced by agro-industrial by-products—on the growth, meat quality, and health parameters of finishing pigs. Eighteen pigs (120 days old, 59.47 ± 0.85 kg) were randomly assigned to three dietary treatments containing 0% (A), 5% (B), or 10% (C) silage. After 60 days, blood samples were collected for hematological and biochemical analyses; this was followed by slaughter for the collection of muscle tissues (triceps brachii, external abdominal oblique) and intestinal digesta (ileum, cecum). Final body weight did not differ among groups (124.54 ± 1.51 kg), and meat composition (fat, protein, collagen, and ash) and pH were unaffected (p > 0.05). ALT levels were significantly lower in group C than in A (p = 0.030). In the ileum, Enterobacteriaceae and Enterococci populations decreased, whereas Lactobacilli increased in B and C (p ≤ 0.05). Cecal Enterococci were reduced in both B and C (p ≤ 0.01), with a tendency for increased Lactobacilli in B. Group C showed higher total phenolic content (p ≤ 0.05) and an improved ω-6/ω-3 ratio (15.09 vs. 17.54 in A). The TBARS values did not differ among treatments. Campylobacter jejuni counts were lower in treatment C, and no Salmonella or Listeria monocytogenes were detected in any samples. The meat color of the triceps brachii was redder in treatments B and C (p = 0.001). Overall, the innovative silage increased phenolic content, supported improvements in gut microbiota, and maintained meat quality, indicating its potential as a sustainable feed ingredient.

Antimicrobial resistance (AMR) emerges and circulates across interconnected human, animal, food, and environmental reservoirs; however, food fermentation systems remain underexplored as indicators of local AMR pressure, even though artisanal dairy fermentations may function as natural sentinels of AMR. In this study, we used an artisanal dairy fermentation chain as a One Health model to investigate whether environmentally exposed lactobacilli can reflect stage-associated shifts in resistance. A total of 1.085 isolates representing 16 Lactobacillus species were recovered from the same artisanal dairy matrix at two fermentation stages: day 5 (“Tyrovolia”; n = 518) and day 30 (“Kopanisti”; n = 567). Susceptibility to 14 antibiotics was evaluated by broth micro-dilution, and L. acidophilus was further screened for selected resistance genes. Overall resistance increased significantly from 69.88% (362/518) at day 5 to 77.25% (438/567) at day 30 (p = 0.0059), while multidrug resistance rose from 37.57% to 60.73% of resistant isolates (p < 0.001). Across the 224 species–antibiotic combinations examined, 129 (57.58%) showed an increased upper MIC limit at day 30, and resistance increased significantly for 9 of the 14 antibiotics tested, with the largest rises observed for metronidazole (RR = 7.67), chloramphenicol (RR = 5.74), quinupristin/dalfopristin (RR = 4.11), vancomycin (RR = 2.78), and trimethoprim (RR = 2.43). In contrast, erythromycin and oxytetracycline resistance declined significantly at the ripened stage. In L. acidophilus, 21 resistance genes were detected in 14/70 day-5 isolates and 19 genes in 13/71 day-30 isolates, but marked genotype–phenotype discordance was observed, including matrix-dependent expression patterns for tetM, ermB, and blaTEM. Collectively, these findings show that environmentally exposed artisanal dairy fermentations can enrich resistance phenotypes and may serve as sensitive sentinels of AMR dynamics at the human–animal–environment interface.

Background/Objectives: Staphylococcus aureus (S. aureus) intramammary infection remains a major global dairy problem due to its contagious nature, its ability to persist and colonize teat/skin and mucosal niches, and the often-limited bacteriological cure achieved with antimicrobial therapy. Beyond udder health, it is relevant to public health because it can enter raw milk chains and serve as a reservoir for antimicrobial resistance determinants that may circulate between dairy animals and humans. Methods: We assessed S. aureus’ ecology in raw ewe milk from 75 sheep farms in Epirus (Greece) by sampling clinically healthy controls (group A) and clinical mastitis cases pre-treatment (group B), followed by resampling at the first post-withdrawal milking after penicillin/streptomycin treatment (group C1—therapeutic protocol 1), oxytetracycline treatment (group C2—therapeutic protocol 2), or enrofloxacin treatment (group C3—therapeutic protocol 3). Results: S. aureus detection was high and comparable across groups (A 23.0%, B 22.0–30.0%, C 20.0–22.0%), and paired analyses showed no significant pre–post shifts in detection/burden within therapeutic protocols (all p > 0.05). Nevertheless, persistence remained evident. The chromosomal gene mecA was detected in S. aureus strains in all groups, ranging from 13.6% in controls to 54.5% post-withdrawal in group C1, and was also present in the pre-treatment group. In paired sampling animals, mecA was mostly stable, with rare emergence or loss. Across antibiotic classes, within-animal resistance transitions were generally uncommon and non-significant (p > 0.05); β-lactam resistance was fully stable (p = 1.00). Descriptively, resistance to protein synthesis inhibitors tended to decline after therapy in protocol 1 and protocol 3, while protocol 3 showed post-treatment gains in fluoroquinolone resistance. By contrast, virulence-associated phenotype traits shifted after therapy: enterotoxigenicity increased post-withdrawal (especially in the C3 group), Staphylococcal Enterotoxin A (SEA) and Staphylococcal Enterotoxin B (SEB) appeared only post-therapy, Staphylococcal Enterotoxin D (SED) increased significantly in paired isolates (p = 0.002), and strong biofilm adherence increased (in C3, p = 1.5 × 10⁻⁵). Conclusions: The detection of S. aureus after therapy suggests that one possibility is that antimicrobial exposure may select for, or otherwise reshape, the residual intramammary population, rather than reliably eliminating it—an outcome that remains clinically relevant for udder health. Moreover, the persistence of mecA/methicillin-resistant Staphylococcus aureus (MRSA)-compatible profiles indicates that milk released to the food chain after withdrawal compliance may still harbor S. aureus with enhanced preservation capacity and significant food safety relevance.